Protocol

Transgenic Cotton

Volume 958 of the series Methods in Molecular Biology pp 211-222

Date:

Agrobacterium-Mediated Transformation of Cotton (Gossypium hirsutum) Shoot Apex with a Fungal Phytase Gene Improves Phosphorus Acquisition

  • Zhiying MaAffiliated withNorth China Key Laboratory for Crop Germplasm Resources of Education Ministry, Agricultural University of Hebei Email author 
  • , Jianfeng LiuAffiliated withCollege of Life Sciences, Hebei University
  • , Xingfen WangAffiliated withNorth China Key Laboratory for Crop Germplasm Resources of Education Ministry, Agricultural University of Hebei

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Abstract

Cotton is an important world economic crop plant. It is considered that cotton is recalcitrant to in vitro proliferation. Somatic embryogenesis and plant regeneration has been successful by using hypocotyl, whereas it is highly genotype dependent. Here, a genotype-independent cotton regeneration protocol from shoot apices is presented. Shoot apices from 3- to 5-day-old seedlings of cotton are infected with an Agrobacterium strain, EHA105, carrying the binary vector pC-KSA contained phytase gene (phyA) and the marker gene neomycin phosphotransferase (NPTII), and directly regenerated as shoots in vitro. Rooted shoots can be obtained within 6–8 weeks. Plants that survived by leaf painting kanamycin (kan) were ­further analyzed by DNA and RNA blottings. The transgenic plants with increased the phosphorus (P) acquisition efficiency were obtained following the transformation method.